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Phage Display: Selecting Straws Instead of a Needle from a Haystack
Department of Pharmaceutical Biology, Faculty of Pharmacy, Aškerčeva 7, Ljubljana, Slovenia
Department of Rheumatology, University Medical Centre Ljubljana, Vodnikova 62, Ljubljana, Slovenia
Department of Biotechnology, Jozef Stefan Institute, Jamova 39, Slovenia
* Author to whom correspondence should be addressed.
Received: 9 December 2010; in revised form: 12 January 2011 / Accepted: 17 January 2011 / Published: 19 January 2011
Abstract: An increasing number of peptides with specific binding affinity to various protein and even non-protein targets are being discovered from phage display libraries. The power of this method lies in its ability to efficiently and rapidly identify ligands with a desired target property from a large population of phage clones displaying diverse surface peptides. However, the search for the needle in the haystack does not always end successfully. False positive results may appear. Thus instead of specific binders phage with no actual affinity toward the target are recovered due to their propagation advantages or binding to other components of the screening system, such as the solid phase, capturing reagents, contaminants in the target sample or blocking agents, rather than the target. Biopanning experiments on different targets performed in our laboratory revealed some previously identified and many new target-unrelated peptide sequences, which have already been frequently described and published, but not yet recognized as target-unrelated. Distinguishing true binders from false positives is an important step toward phage display selections of greater integrity. This article thoroughly reviews and discusses already identified and new target-unrelated peptides and suggests strategies to avoid their isolation.
Keywords: phage display; target-unrelated peptides; decoy
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Vodnik, M.; Zager, U.; Strukelj, B.; Lunder, M. Phage Display: Selecting Straws Instead of a Needle from a Haystack. Molecules 2011, 16, 790-817.
Vodnik M, Zager U, Strukelj B, Lunder M. Phage Display: Selecting Straws Instead of a Needle from a Haystack. Molecules. 2011; 16(1):790-817.
Vodnik, Miha; Zager, Urska; Strukelj, Borut; Lunder, Mojca. 2011. "Phage Display: Selecting Straws Instead of a Needle from a Haystack." Molecules 16, no. 1: 790-817.